title: Production and characterization of a monoclonal antibody against the seed lectin of the Dolichos biflorus plant.
abstract: Spleen cells from mice immunized with the Dolichos biflorus seed lectin were fused with cells from the mouse myeloma Sp2/O-Ag14 cell line to form hybridomas. Those hybridomas producing antibodies against the seed lectin were cloned at least four times and the monoclonal antibodies from clone C11/64-56.28 were characterized and found to be specific for Subunit I of the lectin; they do not react with the structurally similar Subunit II. In previous studies, we have shown that although these two subunits appear to differ only at their COOH-terminal ends, only Subunit I has carbohydrate binding activity. Using a solid phase enzyme immunoassay, the antigenic determinant fr the monoclonal antibody was found to be located on the COOH-terminal cyanogen bromide fragment of this subunit. The monoclonal antibody inhibits the ability of the lectin to agglutinate erythrocytes and N-acetyl-D-galactosamine, the specific hapten for the lectin, inhibits the ability of the antibody to combine with the lectin. These results suggest that the monoclonal antibody recognizes a determinant that is located either at or near the active site of the lectin or that is conformationally interdependent with the active site.
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000917:Antibody Formation; D002459:Cell Fusion; D002460:Cell Line; D002999:Clone Cells; D006822:Hybrid Cells / Q000276:immunology; D007124:Immunoenzyme Techniques; D037102:Lectins; D051379:Mice; D009005:Monosaccharides; D010954:Plasmacytoma; D013154:Spleen / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D037102:Lectins; D009005:Monosaccharides